Anti-Malarial Compositions

ABSTRACT

This disclosure provides antibodies that are useful for preventing and/or treating malaria. The epitope to which the antibodies bind is in close proximity to the conserved proteolytic cleavage site of  P. falciparum  circumsporozoite protein (CSP), and the antibodies provided in this disclosure can prevent cleavage and inhibit  P. falciparum  sporozoites from invading the liver.

This application incorporates by reference the contents of a 11.4 kb text file created on Feb. 25, 2016 and named “00047900245 sequencelisting.txt,” which is the sequence listing for this application.

Each reference cited in this disclosure is incorporated herein by reference in its entirety.

TECHNICAL FIELD

The invention relates to compositions and methods for preventing and treating malaria.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1. Graph of results of biolayer interference assay (BLI) to detect binding of 14 monoclonal antibodies (mAbs) raised against recombinant circumsporozoite protein (“CSP”). Each line represents an individual antibody, and the lines representing antibodies 4B3, 2G9, 3H11, and 5D5 are indicated. See Example 2.

FIGS. 2A-B. Graphs demonstrating binding of anti-CSP mAbs to regions of P. falciparum CSP by ELISA. FIG. 2A shows binding of mAb 5D5 strongly to a long CSP N-terminal region peptide (“N-terminus peptide”). FIG. 2B shows binding of mAb 2G9 to a long CSP repeat region peptide (“Repeat peptide”). See Example 3.

FIG. 3. Photomicrograph of Western Blot showing binding of anti-CSP mAbs 2G9 and 5D5 to P. falciparum sporozoites and rCSP under reducing conditions. Lanes 1 and 3, 25,000 P. falciparum sporozoites; lanes 2 and 4, 0.25 μg rCSP. See Example 4.

FIGS. 4A-C. Mapping of mAb 5D5 to a specific region of the N-terminal CSP domain. FIG. 4A, schematic representation of P. falciparum CSP. FIG. 4B, peptides located in the N terminal region of CSP. Peptide 8, SEQ ID NO:17; peptide 9, SEQ ID NO:18; peptide 10, SEQ ID NO: 19. FIG. 4C, graph of results of peptide mapping by ELISA. See Example 5.

FIG. 5. Graph showing percent inhibition by 1 μg/ml of mAb 5D5 and control mAb 2A10 of parasites in the liver of mice challenged with transgenic P. berghei sporozoites containing a portion of the P. falciparum CSP (repeat region and a small portion of the N-terminal domain, including the cleavage site). See Example 6.

FIGS. 6A-B. Amino acid sequences of the heavy and light chain variable regions of mAb 5D5. CDR sequences are underlined. FIG. 6A, heavy chain variable region (SEQ ID NO:14). FIG. 6B, kappa (light) chain variable region (SEQ ID NO:16). See Example 7.

DETAILED DESCRIPTION

Malaria is an infectious, febrile disease which is caused by protozoa of the genus Plasmodium. Malaria is transmitted by the bites of infected Anopheles mosquitoes and can be caused by any Plasmodium species that infects humans, including, but not limited to, Plasmodium vivax and Plasmodium falciparum.

Circumsporozoite protein (CSP) is the major protein on the surface of the Plasmodium sporozoite. CSP contains an N-terminal domain, a conserved pentapeptide protease cleavage site at the core of region I, a repeat region, a short conserved sequence termed region III, and a C-terminal region with sequence homology to the thrombospondin type-1 repeat superfamily (Doud et al., Proc. Natl. Acad. Sci. USA 109, 7817-22, 2012. CSP is proteolytically processed cysteine protease during invasion of hepatocytes (Coppi et al., J. Exp. Med. 201, 27-33, 2005). Processing occurs on the sporozoite surface when sporozoites contact hepatocytes, resulting in the removal of the amino-terminal third of the protein. The cleavage site is a five amino acid sequence (KLKQP; SEQ ID NO:20), which is highly conserved among P. falciparum isotypes.

This disclosure provides antibodies that bind to an epitope in close proximity to the protease cleavage site of CSP. Binding of the antibodies to this epitope can prevent cleavage and inhibit Plasmodium sporozoites from invading the liver. Because the cleavage site among all species of Plasmodium that infect humans is conserved, the disclosed antibodies can be used to treat or reduce infection in human by all such Plasmodium species.

5D5 Antibodies

Unless otherwise indicated, the term “antibody” means an intact antibody (e.g., an intact monoclonal antibody) or antigen-binding fragment of an antibody (e.g., an antigen-binding fragment of a monoclonal antibody), including intact antibodies or antigen-binding fragments that have been modified or engineered. Modified or engineered antibodies include, but are not limited to, chimeric antibodies, humanized antibodies, and multispecific antibodies (e.g., bispecific antibodies). Examples of antigen-binding fragments include Fab, Fab′, F(ab′)₂, Fv, single chain antibodies (e.g., scFv), and diabodies. Particularly useful subclasses of antibodies include IgG₁, IgG₃, and IgA.

A “5D5 antibody” is an antibody containing the CDR regions of monoclonal antibody (mAb) 5D5, which is described in the specific examples, below. The antibody mAb 5D5 was deposited on Nov. 19, 2013 with The Malaria Research and Reference Reagent Resource Center (MR4), which is managed by BEI Resources, which is managed by ATCC, under Accession No. MRA-1242. Availability of the deposited antibody is not to be construed as a license to use the antibody in contravention of the rights granted under the authority of any government in accordance with its patent laws. mAb 5D5 is a murine IgG₁/κ subclass antibody obtained using the full length recombinant CSP of Plasmodium falciparum (3D7 strain) as an immunogen.

The immunoglobulin heavy chain of mAb 5D5 antibody comprises CDR_(H1) (GYTFTGYGLS; SEQ ID NO:1), CDR_(H2) (IYPRSGNTYYNEKFKGKAT; SEQ ID NO:2), and CDR_(H3) (SWGNSSFVY; SEQ ID NO:3). The immunoglobulin light chain variable region of mAb 5D5 comprises CDR_(L1) (KASQSVTNDVT; SEQ ID NO:4), CDR_(L2) (ASNRYTG; SEQ ID NO:5), and CDR_(L3) (QQDYSSPFT; SEQ ID NO:6). Together, these CDR regions define a binding site for the mAb 5D5 epitope EKLRKPKHKKLK (SEQ ID NO:7).

The locations of the mAb 5D5 epitope (bold and underlined) and the protease cleavage site (bold and italicized) within P. falciparum CSP (SEQ ID NO:8) are shown below:

MMRKLAILSVSSFLFVEALFQEYQCYGSSSNTRVLNELNYDNAGTNL YNELEMNYYGKQENWYSLKKNSRSLGENDDGNNEDN

ADGNPDPNANPNVDPNANPNVDPNANPNVDPNANPNANPNANPN ANPNANPNANPNANPNANPNANPNANPNANPNANPNANPNANPNANP NANPNANPNVDPNANPNANPNANPNANPNANPNANPNANPNANPNAN PNANPNANPNANPNANPNANPNANPNANPNANPNANPNKNNQGNGQG HNMPNDPNRNVDENANANSAVKNNNNEEPSDKHIKEYLNKIQNSLST EWSPCSVTCGNGIQVRIKPGSANKPKDELDYANDIEKKICKMEKCSS VFNVVNSSIGLIMVLSFLFLN

In some variations, a 5D5 antibody is an scFv. Particularly useful 5D5 scFv antibodies are those which comprise an Fc region of the IgG₁, IgG₃, or IgA subclass.

In some variations, the CDR sequences of a 5D5 antibody are interposed between human or humanized immunoglobulin framework regions to reduce or eliminate antigenicity when administered to humans. Methods for humanizing antibodies are known in the art. For example, mouse immunoglobulin constant regions can be replaced with human immunoglobulin constant regions, as described in Morrison et al., Proc. Natl. Acad. Sci. USA 81, 6851-55, 1984; Neuberger et al., Nature 312, 604-08, 1984; U.S. Pat. No. 6,893,625; U.S. Pat. No. 5,500,362; and U.S. Pat. No. 4,816,567.

Alternatively, CDRs can be grafted into human framework regions. The human framework regions can be consensus human framework regions, created by aligning framework regions from several human heavy chain or light chain amino acid sequences to identify a consensus amino acid sequence. Descriptions of CDR grafting are provided, for example, in U.S. Pat. No. 7,022,500; U.S. Pat. No. 6,982,321; U.S. Pat. No. 6,180,370; U.S. Pat. No. 6,054,297; U.S. Pat. No. 5,693,762; U.S. Pat. No. 5,859,205; U.S. Pat. No. 5,693,761; U.S. Pat. No. 5,565,332; U.S. Pat. No. 5,585,089; U.S. Pat. No. 5,530,101; Jones et al., Nature 321, 522-25, 1986; Riechmann et al., Nature 332, 323-27, 1988; Verhoeyen et al., Science 239, 1534-36, 1988; and Winter, FEBS Lett. 430, 92-94, 1998.

Other approaches include SUPERHUMANIZATION™, in which human CDR sequences which are structurally similar to a mouse antibody can be chosen from human germline genes (e.g., U.S. Pat. No. 6,881,557; Tan et al., J. Immunol. 169, 1119-25, 2002); “reshaping,” “hyperchimerization,” or “veneering/resurfacing” (see, e.g., Vaswami et al., Annals of Allergy, Asthma, & Immunol. 81, 105 (1998); Roguska et al., Prot. Engineer. 9, 895-904 (1996); U.S. Pat. No. 5,639,641; U.S. Pat. No. 6,072,035); ACTIVMAB™ technology (Vaccinex, Inc., Rochester, N.Y.), which involves using a vaccinia virus-based vector to express antibodies in mammalian cells, producing high levels of combinatorial diversity of IgG heavy and light chains (e.g., U.S. Pat. No. 6,706,477; U.S. Pat. No. 6,800,442; and U.S. Pat. No. 6,872,518); and HUMAN ENGINEERING™ technology (XOMA (US) LLC (e.g., WO 93/11794; U.S. Pat. No. 5,766,886; U.S. Pat. No. 5,770,196; U.S. Pat. No. 5,821,123; and U.S. Pat. No. 5,869,619).

Production of 5D5 Antibodies

Methods for recombinant production of antibodies are well known in the art. For example, DNA molecules encoding light chain variable regions and heavy chain variable regions can be chemically synthesized using the amino acid sequence information provided in this disclosure. Synthetic DNA molecules can be ligated to other appropriate nucleotide sequences, including, e.g., constant region coding sequences and expression control sequences, to produce conventional gene expression constructs encoding the desired antibody. Production of gene expression constructs is within routine skill in the art. Nucleic acid molecules may comprise coding sequences for one or more of the CDRs of a 5D5 antibody. This disclosure provides the amino acid sequences of the CDRs, and any nucleotide sequence that encodes the desired amino acid sequence may be used to express the desired amino acid sequence. Non-limiting examples of nucleotide sequences include SEQ ID NO:13 (encoding the V_(H) region of mAb 5D5) and SEQ ID NO:15 (encoding the V_(L) region of mAb 5D5).

Nucleic acid molecules can encode one or more of the 5D5 CDRs. In some variations, a nucleic acid molecule encoding one or more of the 5D5 CDR sequences is a cDNA molecule having no introns.

Expression constructs expressing one or more of the 5D5 CDRs can be introduced into host cells through conventional transfection or transformation techniques. Examples of host cells are E. coli cells, Chinese hamster ovary (CHO) cells, HeLa cells, baby hamster kidney (BHK) cells, monkey kidney cells (COS), human hepatocellular carcinoma cells (e.g., Hep G2), and myeloma cells that do not otherwise produce immunoglobulins. Transformed host cells can be grown under conditions that permit the host cells to express the genes that encode the immunoglobulin light or heavy chain variable regions.

Alternatively, amino acid sequences of a 5D5 antibody can be synthesized using a peptide synthesizer, as is known in the art.

Pharmaceutical Compositions and Methods of Use

Pharmaceutical compositions comprising one or more of the 5D5 antibodies disclosed herein typically contain a sufficient concentration of 5D5 antibodies to reduce or prevent cleavage of CSP so that the invasion of the liver by Plasmodium sporozoites is slowed, reduced, or prevented. Pharmaceutical compositions comprise a pharmaceutically acceptable vehicle. Descriptions of suitable pharmaceutically acceptable vehicles, and factors involved in their selection, are found in a variety of readily available sources.

Administration typically is parenteral. Pharmaceutical compositions suitable for parenteral administration comprise various aqueous media such as aqueous dextrose and saline solutions and glycol solutions, and may comprise suitable stabilizing and/or buffering agents. In some variations, 5D5 antibodies are provided in a single or a multi-use vial as a lyophilized sterile powder, under vacuum. Packages or kits containing such vials may also include one or more vials of Bacteriostatic Water for Injection (BWFI), USP, which may contain a preservative (e.g. benzyl alcohol).

A pharmaceutical composition comprising one or more of the 5D5 antibodies disclosed herein can be administered prophylactically to reduce or prevent cleavage of CSP and thereby slow, reduce, or prevent Plasmodium sporozoites from invading the liver. Such compositions can be administered to malaria-naïve individuals (e.g., military personnel, state-department personnel, travelers) entering endemic regions to reduce or prevent infection. In some variations, a pharmaceutical composition comprising a 5D5 antibody can be administered in conjunction with a vaccine composition comprising a malarial antigen such as those disclosed in Mata et al., BioMed Research Int'l. Vol. 2013, Article ID 282913, 2013. Unless otherwise indicated, administration “in conjunction with” a traditional malaria vaccine includes sequential administration (either before or after administration of a traditional malaria vaccine) as well as administration of a 5D5 antibody in a composition comprising an agent that raises an immune response against the Plasmodium.

A pharmaceutical composition comprising 5D5 antibodies can be administered to treat malaria in an individual already infected with a Plasmodium species. In some variations, a pharmaceutical composition comprising 5D5 antibodies is administered in conjunction with one or more other anti-malarial drugs, such as atovaquone and proguanil, chloroquine, doxycycline, mefloquine, and primaquine. Unless otherwise indicated, administration “in conjunction with” an anti-malarial drug includes sequential administration (either before or after administration of an anti-malarial drug) as well as administration of a 5D5 antibody in a composition comprising an anti-malarial drug.

The following examples are provided to illustrate certain particular features and/or embodiments. The examples should not be construed to limit the disclosure to these particular features or embodiments.

Example 1 Antibody Production and Purification

Antibodies were prepared by Precision Antibody, Inc. (Columbia, Md.). Three Balb/c mice were immunized with recombinant full-length P. falciparum CSP (rCSP; SEQ ID NO:8) produced by Pfenex, Inc. (San Diego, Calif.) Serum titers were determined from tail bleed samples via ELISA, and splenocytes were harvested and fused once titers exceeded 1:50,000.

Example 2 Reactivity of Anti-CSP mAbs

A panel of 14 mAbs generated as described in Example 1 were tested for binding to rCSP by biolayer interference assay (BLI), which measures changes in an interference pattern generated from visible light reflected from an optical layer and a biolayer containing a mAb for characterizing its binding profile of rCSP, and by Western blot. The results of the BLI assays are shown in FIG. 1. Most of the antibodies tested demonstrated a strong association to rCSP. Several of the antibodies demonstrated rapid dissociation from rCSP, with 5D5 being the most rapid.

Example 3 Binding of mAbs to CSP Domains

The same panel of 14 mAbs was used in an indirect ELISA to determine binding to long peptides corresponding to the N-terminal (SEQ ID NO:9), repeat (SEQ ID NO:10), and C-terminal (SEQ ID NO:11) domains of CSP. Only mAb 5D5 bound the N-terminal region of CSP (FIG. 2A), while other antibodies bound either to the repeat or C-terminal region of CSP (FIG. 2B).

Example 4 Binding of mAb 5D5 to Native CSP

An immunofluorescence assay (IFA) was used to determine if mAb 5D5 bound to native CSP. Transgenic P. berghei sporozoites containing a portion of the P. falciparum CSP (repeat region and a small portion of the N-terminal domain, including the cleavage site) were the generous gift of Dr. E. Nardin. Sporozoites were obtained by feeding Anopheles stephensi mosquitoes on infected Swiss-Webster mice (Taconic Laboratories) and harvesting sporozoites from dissected salivary glands 18-21 days after the last blood meal. Transgenic P. berghei sporozoites were air-dried onto multispot glass slides and incubated for 1 hour at room temperature with mAbs diluted in PBS+1% BSA. Slides were washed 3× in PBS, incubated for 1 hour at room temperature with a FITC-labeled goat anti-mouse IgG antibody (Kirkegaard and Perry, Gaithersburg, Md.), washed again, and then viewed under a fluorescence microscope.

P. falciparum 3D7 sporozoites air-dried onto multiport glass slides and were incubated for 30 min at 37° C. in a humidified chamber with mAbs diluted in PBS. Slides were washed 3× in PBS and incubated for 30 min at 37° C. in the dark with a FITC-labeled goat anti-mouse IgG antibody (Kirkegaard and Perry, Gaithersburg, Md.) diluted 1:40 in PBS containing 0.02% Evans blue. Slides were washed again, mounted with VECTASHIELD® mounting media, and viewed under an epifluorescence microscope.

A weak fluorescence signal was detected with mAb 5D5 compared with the signal generated by control mAb 2G9, which recognizes the CSP repeat region.

An IFA was also performed using a chimeric rodent malaria parasite P. berghei that expresses a larger portion of the N-terminal region (amino acids 22-92) of the P. falciparum CSP. Again, mAb 5D5 generated a weak fluorescence signal as compared to mAb 2G9. The mAb 5D5 recognized P. falciparum 3D7 parasites in situ but required a higher concentration compared to mAb 2G9 for a signal to be detected.

Binding to native CSP also was examined via western blot with P. falciparum 3D7 sporozoite lysate and rCSP under reducing conditions (FIG. 3). Differential binding was seen with the mAb 5D5, whereas the control mAb 2G9 bound to rCSP in sporozoite lysate and to rCSP.

Example 5 Peptide Mapping of Anti-CSP mAbs

To define further which area of the N-terminal region mAb 5D5 recognizes, an indirect ELISA was performed with overlapping 15-mer peptides spanning the entire CSP. This mapping was conducted by AscentGene, Rockville, Md., using peptides diluted in DMSO to a concentration of 2 μg/ml. Plates were blocked with BSA and incubated with 1.3 μg/well of each mAb. An HRP-conjugated anti-mouse secondary antibody (1:10,000) was used, followed by detection with TMB substrate.

The results are shown in FIG. 4C. Strong reactivity of the 5D5 mAb was found only to peptide 9 (SEQ ID NO:18), which consisted of amino acids 81-95 of CSP. No reactivity was seen with mAb 5D5 to peptide 8 (amino acids 71-85 of CSP; SEQ ID NO:17) or peptide 10 (amino acids 91-106 of CSP; SEQ ID NO:19). The putative 5D5 epitope was identified as EDNEKLRKPKHKKLK (SEQ ID NO:7).

To confirm the epitope, competitive ELISA assays were performed using a series of 9-mer peptides (peptide A, SEQ ID NO:21; peptide B, SEQ ID NO:22; peptide C, SEQ ID NO:23; peptide D, SEQ ID NO:24; peptide E, SEQ ID NO:25) encompassing the sequence corresponding to peptides 8-10 to identify which peptides competed with rCSP for binding to mAb 5D5. The results are shown in Table 1. The results confirm that peptide 9 competes with rCSP for binding to mAb 5D5.

TABLE 1 Peptide concen- tration A B C D E 8 9 10 rCSP 0.01 3.570 3.510 3.488 3.453 3.392 3.353 0.246 3.336 0.340 μg/ml 0.001 1.052 1.026 0.976 0.989 0.925 0.952 0.190 0.920 0.191 μg/ml NC 0.245 0.204 0.230 0.226 0.211 0.219 0.229 0.211 0.198 PC 3.595 3.405 3.557 3.506 3.471 3.417 3.442 3.393 3.490 0.01 μg/ml PC 1.352 1.315 1.311 1.304 1.228 1.212 1.208 1.168 1.174 0.001 μg/ml

Example 6 Passive Transfer of Anti-CSP mAb Decreases Parasite Liver Load

Challenge studies were performed with C57Bl/6 mice and live chimeric P. berghei sporozoites expressing either the repeat region (and a small portion of the N-terminal domain) or the majority of the N terminal domain of P. falciparum, both of which contain the cleavage site and the 5D5 epitope. Each mouse was injected intravenously with 300 μg of mAb 5D5 or one of the positive control mAbs 3D11 or 2A10 in PBS just before challenge with 10,000 sporozoites suspended in 100 μl of PBS containing 1% normal mouse serum. Mice were euthanized 40-42 hours post challenge, and their livers were excised. Total RNA was extracted from the livers and used to estimate liver parasite load by real time PCR as described in Bruna-Romero et al., Int J. Parasitol. 31, 1499, 2001.

The results are shown in FIG. 5. Liver load of parasites in mice to which mAb 5D5 was administered was significantly decreased compared to the naïve control group.

Example 7 Analysis of mAb 5D5 Heavy and Light Chains

Total RNA Extraction.

Total RNA was extracted from hybridoma cells using Trizol Reagent (Invitrogen Catalog Number 15596) and prepared according to the manufacturer's protocol. Isopropanol-precipitated RNA was resuspended in sterile RNAse/DNAse free H₂O and the absorbance at A260 determined. Electrophoresis on a 1% TAE-agarose gel was used to determine quality.

First-Round RT-PCR.

5 μg of RNA was used for the first strand cDNA preparation using SUPERSCRIPT® III (Invitrogen catalog #18080-051). Oligo-dT was used as primer. cDNA was purified using Qiagen QIAQUICK® columns and tailed using terminal deoxyribonucleotide transferase (Invitrogen catalog #10533-065). The polyadenylated first strand cDNA was purified using Qiagen QIAQUICK® columns as before. PCR was performed with reverse primers specific for the heavy and light chains and with a common oligodT primer as a forward primer. Reverse primers were located in the constant regions of heavy and light chains. No restriction sites were engineered into the primers.

PCR Conditions:

i. Forward primer only: 50° C., 5 cycles ii. Add reverse primer(s) iii. Denaturation step 94° C., 120 seconds iv. 35 Cycles of: 94° C., 30 seconds 55° C., 30 seconds 72° C., 120 seconds

Analysis of PCR Results.

PCR reaction samples were analyzed on a preparative TAE-1% agarose gel to visualize the amplified DNA fragments. The correct antibody variable region DNA fragments should have a size between 600-750 base pairs. The amplified PCR products are excised from the gel and purified using Qiagen QIAQUICK® columns.

The purified PCR products were ligated into EcoRV-digested pBluescript using a QUICK-LIGATION® kit (New England BioLabs, Inc. Catalog #M2200S) and transformed into E. coli strain DH5-α on X-Gal/IPTG LB plates. The next day, white colonies were placed into 3 ml of LB+amp broth, grown overnight, and purified using Qiagen minicolumns, followed by restriction digestions and gel electrophoresis. A minimum of 6 correctly sized clones of each light and heavy chain were sequenced.

Sequencing.

The cloned DNAs were sequenced by Sequetech (Mountain View, Calif.) using an ABI 3730XL Capillary Sequencer using primers which flank both ends of the cDNA insert. The resulting DNA files were assembled and edited using VECTOR NTI®. The resulting assemblies translated to provide a deduced amino acid sequence.

The heavy chain insert encodes an IgG1 isotype murine antibody. Full-length clones included a 51 bp 5′ untranslated leader preceding a canonical 19 amino acid signal sequence. The light chain insert encodes an kappa isotype murine antibody. Full-length clones included a 27 bp 5′ untranslated leader preceding a canonical 20 amino acid signal sequence.

The amino acid sequences of the heavy and light chains are shown in FIG. 6A and FIG. 6B. The CDRs of mAb 5D5 were identified using the Kabat database (bioinf<dot>org<dot>uk<forward slash>abs<forward slash>#cdrdef); see “Protein Sequence and Structure Analysis of Antibody Variable Domains” in Antibody Engineering Lab Manual, Dübel & Kontermann, eds., Springer-Verlag, Heidelberg, 2001). 

1. A cDNA molecule, comprising a coding sequence for a complementarity determining region of a heavy chain (CDR_(H)) of an antibody, wherein the CDR_(H) is selected from the group consisting of: (i) a first CDR_(H) comprising the amino acid sequence SEQ ID NO:1; (ii) a second CDR_(H) comprising the amino acid sequence SEQ ID NO:2; (iii) a third CDR_(H) comprising the amino acid sequence SEQ ID NO:3; and (iv) at least one CDR_(H) of the antibody deposited under Accession No. MRA-1242.
 2. The cDNA molecule of claim 1, which comprises: (i) a coding sequence for the first CDR_(H); (ii) a coding sequence for the second CDR_(H); (iii) a coding sequence for the third CDR_(H); (iv) a coding sequence for the first CDR_(H) and a coding sequence for the second CDR_(H); (v) a coding sequence for the first CDR_(H) and a coding sequence for the third CDR_(H); (vi) a coding sequence for the second CDR_(H) and the coding sequence for a third CDR_(H); or (vii) a coding sequence for the first CDR_(H), a coding sequence for the second CDR_(H), and a coding sequence for the third CDR_(H).
 3. The cDNA molecule of claim 1, which comprises a coding sequence for the at least one CDR_(H) of the antibody deposited under Accession No. MRA-1242.
 4. A cDNA molecule, comprising a coding sequence for a complementarity determining region of a light chain (CDR_(L)) of an antibody, wherein the CDR_(L) is selected from the group consisting of: (i) a first CDR_(L) comprising the amino acid sequence SEQ ID NO:4; (ii) a second CDR_(L) comprising the amino acid sequence SEQ ID NO:5; (iii) a third CDR_(L) comprising the amino acid sequence SEQ ID NO:6; and (iv) at least one CDR_(L) of the antibody deposited under Accession No. MRA-1242.
 5. The cDNA molecule of claim 4, which comprises: (i) a coding sequence for the first CDR_(L); (ii) a coding sequence for the second CDR_(L); (iii) a coding sequence for the third CDR_(L); (iv) a coding sequence for the first CDR_(L) and a coding sequence for the second CDR_(L); (v) a coding sequence for the first CDR_(L) and a coding sequence for the third CDR_(L); (vi) a coding sequence for the second CDR_(L) and the coding sequence for a third CDR_(L); or (vii) a coding sequence for the first CDR_(L), a coding sequence for the second CDR_(L), and a coding sequence for the third CDR_(L).
 6. The cDNA molecule of claim 4, which comprises a coding sequence for the at least one CDR_(L) of the antibody deposited under Accession No. MRA-1242.
 7. The cDNA molecule of claim 3, further comprising a coding sequence for the at least one CDR_(L) of the antibody deposited under Accession No. MRA-1242.
 8. A method of producing an antibody, comprising expressing a cDNA molecule selected from the group consisting of: a) a cDNA molecule which comprises a coding sequence for a complementarity determining region of a heavy chain (CDR_(H)) of an antibody, wherein the CDR_(H) is selected from the group consisting of (i) a first CDR_(H) comprising the amino acid sequence SEQ ID NO:1; (ii) a second CDR_(H) comprising the amino acid sequence SEQ ID NO:2; (iii) a third CDR_(H) comprising the amino acid sequence SEQ ID NO:3; and (iv) at least one CDR_(H) of the antibody deposited under Accession No. MRA-1242; b) a cDNA molecule which comprises (i) a coding sequence for the first CDR_(H); (ii) a cDNA molecule which comprises a coding sequence for the second CDR_(H); (iii) a cDNA molecule which comprises a coding sequence for the third CDR_(H); (iv) a cDNA molecule which comprises a coding sequence for the first CDR_(H) and a coding sequence for the second CDR_(H); (v) a cDNA molecule which comprises a coding sequence for the first CDR_(H) and a coding sequence for the third CDR_(H); (vi) a cDNA molecule which comprises a coding sequence for the second CDR_(H) and the coding sequence for a third CDR_(H); or (vii) a cDNA molecule which comprises a coding sequence for the first CDR_(H), a coding sequence for the second CDR_(H), and a coding sequence for the third CDR_(H); c) a cDNA molecule which comprises a coding sequence for the at least one CDR_(H) of the antibody deposited under Accession No. MRA-1242; d) a cDNA molecule which comprises a coding sequence for a complementarity determining region of a light chain (CDR_(L)) of an antibody, wherein the CDR_(L) is selected from the group consisting of (i) a first CDR_(L) comprising the amino acid sequence SEQ ID NO:4; (ii) a second CDR_(L) comprising the amino acid sequence SEQ ID NO:5; (iii) a third CDR_(L) comprising the amino acid sequence SEQ ID NO:3; and (iv) at least one CDR_(L) of the antibody deposited under Accession No. MRA-1242; e) a cDNA molecule which comprises (i) a coding sequence for the first CDR_(L); (ii) a cDNA molecule which comprises a coding sequence for the second CDR_(L); (iii) a cDNA molecule which comprises a coding sequence for the third CDR_(L); (iv) a cDNA molecule which comprises a coding sequence for the first CDR_(L) and a coding sequence for the second CDR_(L); (v) a cDNA molecule which comprises a coding sequence for the first CDR_(L) and a coding sequence for the third CDR_(L); (vi) a cDNA molecule which comprises a coding sequence for the second CDR_(L) and the coding sequence for a third CDR_(L); or (vii) a cDNA molecule which comprises a coding sequence for the first CDR_(L), a coding sequence for the second CDR_(L), and a coding sequence for the third CDR_(L); f) a cDNA molecule which comprises a coding sequence for the at least one CDR_(L) of the antibody deposited under Accession No. MRA-1242; and g) a cDNA molecule which comprises a coding sequence for the at least one CDR_(L) of the antibody deposited under Accession No. MRA-1242 and a coding sequence for the at least one CDR_(H) of the antibody deposited under Accession No. MRA-1242.
 9. The method of claim 8, wherein the cDNA molecule comprises a coding sequence for the variable region of the heavy chain of the antibody deposited under Accession No. MRA-1242.
 10. The method of claim 8, wherein the cDNA molecule comprises a coding sequence for the variable region of the light chain of the antibody deposited under Accession No. MRA-1242.
 11. The method of claim 9, wherein the cDNA molecule further comprises a coding sequence for the variable region of the light chain of the antibody deposited under Accession No. MRA-1242. 